Abstract
Many biological experiments can be successfully carried out on tissues isolated under well-defined experimental conditions, but for some physiological or pharmacological studies, knowledge of the in vivo elemental composition and/or structure is essential. We have developed a method for rapid freezing of organs in situ, with a new spring-loaded, hand held, clamping device that holds two apposing melting Freon popsicles (Fig. 5). The melting Freon surfaces that contact the tissue are at the melting point of Freon 22, and these mold to the unevenness of the tissue surface. Rapidly frozen samples of liver, pancreas or mesentery were obtained by placing an anesthetized rat in a cloth sling over an environmental chamber kept at 37°C, 100% humidity (Fig. 5). A small incision is made in the abdomen, and the abdominal wall is manipulated until a lobe of liver protrudes through the incision, while taking care not to touch the surface of the organ. Immediately, the Freon clamper is activated to snap freeze the tissue.
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More From: Proceedings, annual meeting, Electron Microscopy Society of America
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